Fig. 5.

Analysis of lipid abundance and lipid biosynthesis in NIH-3T3 cells transduced with ATF6α(1-373). (A) NIH-3T3 cells were transduced with empty vector (black bars) or ATF6α(1-373) (gray bars) retroviruses and assessed at 48 hours post-transduction. Total amounts of phosphatidylcholine (PtdCho), phosphatidylethanolamine (PtdEtn), cholesterol (Chol), sphingolipid (Sphing) and cholesterol ester (Chol ester) were determined by flame ionization and normalized to total cellular protein. The results are the mean ± s.d. of triplicate determinations and are representative of two independent experiments. (B,C) NIH-3T3 cells were transduced with the indicated retroviruses. At 48 hours post-transduction, cells were metabolically labeled with [¹⁴C]acetate for 2 hours. The incorporation of radiolabel into total fatty acids (B) and PtdCho (C) was determined as described in Materials and Methods and normalized to 10⁷ cells. The data are the mean ± s.d. of triplicate determinations and are representative of two independent experiments.